Graft copolymer for cation- exchange chromatography

Abstract

The invention relates to chromatographic separating materials having improved binding capacity for biological constituents in cell culture supernatants, or animal or human body fluids, in particular for monoclonal antibodies. The present invention likewise relates to the preparation of separating materials of this type, and to the use thereof, in particular for the removal of charged biopolymers from corresponding liquids.

Description

[0001]The invention relates to a separating material having improved binding capacity, to the preparation thereof, and to the use thereof for the removal of charged biopolymers from liquids.PRIOR ART[0002]Chromatography is one of the most suitable methods for the isolation of proteins. Monoclonal antibodies can be purified, for example, by affinity chromatography using protein A ligands. Binding to the ligand from the cell culture supernatant is possible without adaptation of pH and salt concentration. Nevertheless, these sorbents can only be employed to a limited extent due to their high costs and due to bleeding-out of the ligand.[0003]The use of high-capacity ion exchanger resins is a favourable alternative. However, the conduction value in the cell culture supernatant must be reduced in order that binding to the ion exchanger takes place. This can be carried out by desalination or by dilution of the supernatant. Both possibilities are undesired, in particular, in the case of lar...

AI Technical Summary

Benefits of technology

[0221]The invention likewise relates to a kit for the purification or separation of biopolymers from one or more other substances in a liquid. The kit consists of a chromatography column which is packed with the separating material according to the invention, one or more buffers and a pack leaflet with written instructions. The liquid is adjusted to a pH of, for example, 5.5 using a buffer and brought into contact with the chromatography column. The column is firstly washed with a wash buffer, giving one fraction of the non-binding constituents, and the biopolymers are then desorbed using an elution buffer of higher ion strength, for example using 1 M NaCl solution, and obtained in a second fraction.

[0222]The present description enables the person skilled in the art to apply the invention comprehensively. Even without further comments, it is therefore assumed that a person skilled in the art will be able to utilise the above description in the broadest scope.

[0223]If anything is unclear, it goes without saying that the publications and patent literature cited should be consulted. Accordingly, these documents are regarded as part of the disclosure cont

Problems solved by technology

Nevertheless, these sorbents can only be employed to a limited extent due to their high costs and due to bleeding-out of the ligand.

Both possibilities are undesired, in particular, in the case of large-volume production processes.

However, there are only a few commercially available adsorbents which are suitable for binding proteins and in particular antibodies from cell culture supernatants.

However, this gel was not developed especially for antibodies and binds 45 mg/ml of BSA.

In addition, there are further commercially available products containing synthetic ligands which are only able to bind antibodies from buffer solutions.

On use for the treatment of cell culture supernatants, unsatisfactory or no binding are obtained.

This result is probably caused by components in the cell culture supernatant which have an interfering behaviour.

However, no separating materials are known to date which can be prepared by a process which is simple to carry out using inexpensive starting materials and which have such high separation activities, in particular wit

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